Methods for Inhibiting the Development of Huntington&#39;s Disease

ABSTRACT

This invention relates to Huntington&#39;s disease and more specifically to methods for inhibiting the development of or treating Huntington&#39;s disease by administering estrogen, testosterone, precursors thereof or combinations thereof.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/334,643, filed Jul. 17, 2014, which is a continuation of U.S.application Ser. No. 12/961,250, filed Dec. 6, 2010, now U.S. Pat. No.8,895,541, which is a continuation-in-part of U.S. application Ser. No.12/719,361, filed Mar. 8, 2010, which is a continuation of U.S.application Ser. No. 11/627,978, filed Jan. 28, 2007, now U.S. Pat. No.7,674,592, which is a continuation of U.S. application Ser. No.10/654,850, filed Sep. 4, 2003, now abandoned, which claims the benefitof U.S. Provisional Application No. 60/443,397, filed Jan. 29, 2003 andU.S. Provisional Application No. 60/408,184, filed Sep. 4, 2002.

The entire teachings of the above patent and applications areincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to Huntington's Disease (HD) andtrinucleotide repeat disorders. The present invention further relates tomethods for treating individuals having HD, or reducing HD associatedsymptoms.

BACKGROUND OF THE INVENTION

Huntington's disease is a degenerative, neurological disease that isalmost exclusively inherited from a parent. It is believed that about30,000 people in the United States currently have Huntington's disease(HD) and about 150,000 are at risk of having inherited the disease froma parent. Individuals who are at risk of developing Huntington's diseaseinherit a mutated HD gene from a parent that codes for a protein nowknown as a mutated huntingtin protein. The HD gene is located onchromosome 4 and is characterized by an expanded trinucleotide repeatmade up of cytosine, adenine and guanine (CAG). HD is an autosomaldominant disorder. Specifically the HD gene is located on anonsex-linked chromosome, which means that men and women are equally atrisk of inheriting the HD gene and, if the gene is inherited from justone of either parent, the inheriting individual will inevitably developthe disease.

HD is generally difficult to treat and individuals having HD suffersymptoms including brain tissue loss, weight loss, decreased cognitiveability loss of motor function, and several others. Physicians haveoften struggled to control and minimize these symptoms.

Accordingly, a need exists for a better understanding of HD. A furtherneed exists for an effective treatment for HD as well as methods andcompositions to reduce the symptoms associated with HD.

SUMMARY OF THE INVENTION

The present invention relates to methods of treating an individualhaving Huntington's Disease (HD) or a mutant HD gene having greater thanor equal to about 36 CAG repeats (e.g., between about 36 and about 120CAG repeats). The method involves selecting an individual having HD oran HD gene having greater than or equal to about 36 CAG repeats; andadministering an amount of estrogen, testosterone, a precursor thereof,or a combination thereof. In an embodiment, estrogen is administered toa female in an amount that ranges from about 0.025 mg to about 10 mg,and testosterone is administered to a male in an amount that ranges fromabout 1 mg to about 35 g. The “estrogen/testosterone compound” asreferred to herein is intended to mean “estrogen, a precursor thereof,or a combination thereof” when administered to a female individual and“testosterone, a precursor thereof, or a combination thereof” whenadministered to a male individual. The method can further include, in anaspect, measuring levels of estrogen, testosterone, a precursor thereof,or a combination thereof in the individual e.g., before administration.In this case, the amount of the administration of theestrogen/testosterone compound is sufficient to maintain certain levelsof the compound in individuals with HD, especially in cases in which thelevels decrease as the disease progresses. The levels to be maintainedare those that are homeostatic levels of estrogen or testosterone inindividuals normally associated with a 20-25 year old population. Inwomen, a homeostatic level of an average amount of estrogen in a 20-25year old female ranges between about 100 pg/ml and about 185 pg/ml.Homeostatic levels in male individuals of free testosterone is betweenabout 20 pg/ml and about 40 pg/ml, and total testosterone is between 300ng/dL and about 900 ng/dL. The method of treatment further includesreducing one or more symptoms of HD e.g., weight loss, loss of fine orgross motor function, a loss in cognitive function, chorea, loss ofbrain tissue or a combination thereof. One or more symptoms are reducede.g., by a range between about 5% and about 100%. The methods of thepresent invention includes assessing the presence or absence of a mutantHD gene having between about 36 and 120 CAG repeats (e.g., using a DNAtest, PCR testing, as further described herein).

The present invention also pertains to methods for reducing one or moresymptoms associated with HD in an individual, wherein the methodincludes selecting an individual having HD; and administering an amountof estrogen, precursors thereof, or a combination thereof to a female,wherein the amount ranges from about 0.025 mg to about 10 mg oradministering an amount of testosterone, precursors thereof, or acombination thereof to a male, wherein the amount ranges from about 1 mgto about 35 g; wherein one or more symptoms associated with HD in theindividual is reduced, as compared to an individual with HD notsubjected to the estrogen/testosterone compound.

The present invention further embodies preventing weight loss, orreducing the amount of weight loss in an individual having HD, byselecting an individual having HD; and administering an amount ofestrogen to a female, precursors thereof, or a combination thereof,wherein the amount of estrogen ranges from about 0.025 mg to about 10mg, and administering an amount of testosterone to a male, wherein theamount of testosterone ranges from about 1 mg to about 35 g. Suchadministration prevents weight loss or reduces the amount of weight lossin the individual with HD, as compared to an individual with HD notsubjected to such administration. In an embodiment, the amount of weightloss is reduced by between about 5% and about 100%. In an aspect, themethods of the present invention involves selecting an individual havingHD by assessing an individual for the presence or absence of a mutanthuntingtin protein having a polyglutamine tract, or mutant HD genehaving a CAG repeat.

Yet the present invention further relates to methods for treating anindividual having HD, by assessing an individual for the presence orabsence of HD or the presence or absence of a HD gene having betweenabout 36 CAG repeats and about 120 CAG repeats; assessing one or morelevels of estrogen, testosterone, precursors thereof, or a combinationthere in the individual; and administering an amount of estrogen,testosterone, or a combination thereof to the individual to maintainestrogen at an average level in a female individual between about 100pg/ml and about 185 pg/ml; or in a male individual to maintain a levelof free testosterone is between about 20 pg/ml and about 40 pg/ml,and/or of total testosterone is between 300 ng/dL and about 900 ng/dL.The estrogen, testosterone, or a combination thereof can beadministered, for example, periodically, daily, or weekly.

The present invention also pertains to methods of treating an individualhaving a trinucleotide repeat disorder by selecting an individual havinga trinucleotide repeat disorder; and administering to a femaleindividual an amount of estrogen, precursors thereof or a combinationthereof; or administering to a male individual an amount oftestosterone, precursors thereof or a combination thereof; wherein theindividual having a trinucleotide repeat disorder is treated. Examplesof trinucleotide repeat disorder includes HD, Dentatorubropallidoluysianatrophy, Kennedy disease, Spinocerebellar ataxia (SA) Type 1, SA Type 2,SA Type 3, SA Type 6, SA Type 7, and SA Type 17.

Additionally, methods of inhibiting a mutant huntingtin protein in anindividual having HD is also encompassed the present invention. Suchmethods include administering to a female individual an amount ofestrogen, precursors thereof or a combination thereof; or administeringto a male individual an amount of testosterone, precursors thereof or acombination thereof; wherein the mutant huntingtin protein is inhibited.In an embodiment, estrogen binds the mutant huntingtin protein andinhibits its function.

Furthermore, a method of the present invention for inhibiting thedevelopment of Huntington's disease in an individual who is at risk ofdeveloping the disease, includes some or a combination of the followingsteps: determining that the individual exhibits a trinucleotide repeatpattern, consisting of cytosine, adenine, and guanine, is of sufficientnumber to indicate a risk for developing HD; establishing that a serumlevel of a preselected hormone in the individual is below normal;administering one or more hormones, selected from a group consisting ofestrogen, testosterone, their respective precursors, and esters ofestrogen, testosterone, and their respective precursors, in amountssufficient to inhibit development of the disease (e.g., at a level ofestrogen in women or testosterone in men comparable to that of a 20-25year old population), wherein the individual, in an embodiment, exhibitsan expanded trinucleotide repeat pattern greater than 36. In anembodiment, the individual exhibits repeats higher than 38 includingexpanded trinucleotide repeat patterns equal to or greater than 43 oreven equal to or greater than 63. In an aspect, the individual exhibitsa mutated huntingtin polyglutamine protein comprising greater than 38glutamines. To establish that an individual's serum level is belownormal, the method includes, in an embodiment, the step of testing theindividual's blood sample by a suitable method such as a polymerasechain reaction.

The method of the present invention further includes, in an embodiment,the step of predetermining the rate at which one or more of the hormonesbinds to a polyglutamine located at an end of said huntingtinpolyglutamine protein to determine a time to begin said administeringstep and said sufficient amount of said one or more hormones, whereinsaid predetermining step comprises the steps of, obtaining one or moresamples of a huntingtin polyglutamine protein with known numbers ofglutamines; mixing said sample with a labeled estradiol source and abuffering solution; measuring the binding affinity of the labeledestradiol source to the huntingtin polyglutamine protein. In an aspect,the binding affinity is preferably measured with a gamma counter and isequal to or less than about 50,000 counts per minute.

Another method for determining a time for administering a hormonetreatment to inhibit the development of Huntington's disease in anindividual who is at risk of developing the disease, generally includesthe steps of: determining a plurality of binding affinities of estradiolto a mutant huntingtin polyglutamine protein with known numbers ofglutamines; and measuring the serum level of hormone in said individualto determine if said serum level is below normal. In an aspect, theaffinity can be measured with a gamma counter and typically is equal toor less than about 50,000 counts per minute and may be equal to or lessthan about 40,000 counts per minute. The mixing step may furthercomprise mixing said labeled hormone source with a buffering solution.

The present invention has numerous advantages. The present inventionprovides for an effective treatment for HD. Particularly, the methods ofthe present invention allow for a reduction in symptoms associated withHD. Unexplained weight loss is a tell-tale sign of HD and generally getsmore pronounced as the disease progresses. For example, most HD patientsconsume approximately 2,000-4,000 calories/day, which is more than twicethe recommended amount and still lose weight. The methods of the presentinvention surprisingly and significantly reduce this remarkable weightloss experienced by an individual with HD.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the amount of time (in seconds) animals fromgroups E7, E12, E26, C16, C3 and C4 were able to traverse a ½ in beam ata certain time (in weeks).

FIG. 2A is a graph showing the amount of time (in seconds) animals from{grave over (g)}roups E7 and C4 were able to traverse a ½ in beam at acertain time (in weeks).

FIG. 2B is a bar graph showing the extent (0=no movement, 5=completedtask, (transverse the entire beam) animals from groups E7 and C4 wereable to traverse a ½ in beam at a certain time (in weeks).

FIG. 3A is a graph showing the amount of time (in seconds) animals fromgroups E12 and C3 were able to traverse a ½ in beam at a certain time(in weeks).

FIG. 3B is a bar graph showing the extent (0=no movement, 5=completedtask, (transverse the entire beam) animals from groups E12 and C3 wereable to traverse a ½ in beam at a certain time (in weeks).

FIG. 4A is a graph showing the amount of time (in seconds) animals fromgroups E26 and C16 were able to traverse a ½ in beam at a certain time(in weeks).

FIG. 4B is a bar graph showing the extent (0=no movement, 5=completedtask, (transverse the entire beam) animals from groups E26 and C16 wereable to traverse a ½ in beam at a certain time (in weeks).

DETAILED DESCRIPTION OF THE INVENTION

A description of preferred embodiments of the invention follows.

The present invention relates to methods for testing for and inhibitingthe development of Huntington's disease (HD) in individuals byadministering an amount of estrogen (e.g., Estradiol), testosterone, aprecursor thereof, or a combination thereof (referred hereincollectively as “estrogen/testosterone administration”). In particular,the present invention relates to treating and/or preventing HD in anindividual by estrogen/testosterone administration. The“estrogen/testosterone compound” as referred to herein is intended tomean “estrogen, a precursor thereof, or a combination thereof” whenadministered to a female individual and “testosterone, a precursorthereof, or a combination thereof” when administered to a maleindividual. To treat an individual with HD disease or a trinucleotiderepeat disorder means to alleviate, ameliorate or reduce the severity ofone or more of HD symptoms. Prevention of HD or trinucleotide repeatdisorder refers to delaying or suppressing the onset of the one or moresymptoms of HD. Additionally, the present invention encompasses reducingthe severity of one or more HD symptoms, which refers to minimizing theextent of one or more HD symptoms that are experienced by theindividual.

In an embodiment, the methods of the present invention utilize balancedhormone treatment, including estrogen, testosterone, their precursorssuch as DHEA and progesterone; and their esters to inhibit and/or treatHD. Accordingly, the present invention involves assessing the levels ofestrogen, testosterone, precursors thereof and/or a combination thereof,and administering an amount of estrogen (e.g., Estradiol), testosterone,precursors thereof, or a combination thereof to bring one or more levelsto a desired amount or within a normal range, as further describedherein.

As demonstrated by the data in the Exemplification section, one of themost significant symptoms of HD in humans is the continued loss of bodyweight even at extreme caloric intake. In generally, individualssuffering from HD routinely continue to lose weight as the diseaseprogresses even at 4000-5000 calorie/day intake. Along with the extremeweight loss, brain and motor function abnormalities occur, as furtherdescribed herein. Individuals with HD have cognitive function, motorfunction and gait that are extremely impaired. Additionally HDindividuals are often associated as having mood swings and outburst ofrage.

Generally, HD is characterized by a trinucleotide repeat or a CAG repeatthat is also further described herein. An individual having HD typicallyhas greater than or equal to 36 CAG repeats. The mouse model used in thedata of the Exemplification utilizes severely diseased mice that have anaverage of about 159.6 CAG repeats and demonstrate the sameabnormalities as individuals having HD. Extreme weight loss duringprogression of the disease was observed. Cognitive inabilities indecision making, and tremendous hesitation (obsessive compulsivebehavior) as well as elliptic seizure occurred in non-treated mice(i.e., the control). Some aggressive cage behavior was observed with thecontrol animals. The data show that administration of either estrogenand or testosterone, surprisingly, abolished or greatly reduced thesesymptoms in severely affected HD mice. As the disease state progressed,the untreated animals displayed symptomatic abnormalities, includingextreme weight loss (up to 30% Males, 28% Females). Untreated mice werenot able to perform certain cognitive and motor functions such asdeciding to cross a beam (impaired cognition) and demonstrated inabilityto traverse a beam to a safety platform once placed on it. The treatedanimal showed a remarkable improvement over the control animals in theirability to transverse the ½ inch beam in the allotted time frame. Theydemonstrated cognitive ability and showed little hesitation in decidingto cross. Once moving along the beam, the treated animals were able tocontrol their forelimb and hind limb coordination to show little or noimpairment. Cognitive and motor ability in the treated mice continuedfor a significant amount of time after the untreated mice were unable totraverse the beam. As described in the Exemplification, treated micetraversed the beam two to two and a half weeks after the control animalswere unable to move on the beam (a 16.6% improvement over the timecourse of the study).

The data of the Exemplification also shows that the treated animalsshowed a marked improvement in weight retention, a symptom that isstrongly associated with HD progression. Aziz, N. Ahmad, et al., JON3062: 1872-1878 (January 2009). Also, transgenic for severe disease, thetreated mice's overall weight loss was only a fraction of the untreatedanimal, e.g., at as little as 11% for males and as little as 9% forfemales. As compared to untreated mice, a significant improvement inweight loss was seen.

Taken together, these data show that the administration of estrogen andor testosterone is protective against Huntington's disease and symptomsassociated with the disease. This protection is effective at more thanone dose level and is shown to be quantitative in at least four areas ofmeasurement: (1) weight loss, (2) cognitive function, (3) motor functioncoordination and (4) brain weight assessment. The present inventionprovides an effective therapy for HD.

In general, HD is a genetic, neurodegenerative disease and affectsmuscle coordination and certain cognitive functions. HD ischaracterized, in part, by neural degeneration specific to the spinyneuron in the striatum. The disease, which is characterized by anexpanded CAG (cytosine-adenine-guanine), a polyglutamine repeat, oftenmanifests itself with aging and is associated with the onset of HD. TheHD gene, also referred to as the HTT gene, is located on the short armof chromosome 4 at 4p16.3 and codes for huntingtin protein. CAG is thegenetic code for the amino acid, glutamine, so a series of them resultsin the production of a chain of glutamine known as a polyglutamine tract(or polyQ tract). The mutant form of the huntingtin protein having anexpanded polyQ tract greater than 36Q causes HD. The present inventioninvolves individuals having HD and/or those having an HD gene withgreater than or equal to about 36 CAG repeats (e.g., about 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, or 120 CAG repeats).

Symptoms of HD include psychomotor related symptoms, such as chorea,which refers to jerky, random, or uncontrollable movements. As thedisorder progresses, psychomotor symptoms also include rigidity,writhing motions or abnormal posturing. Often, assessment of motorfunction can be done by observing the individual's walk, posture,movement and/or gait. One of skill in the art or a healthcare providercan assess such HD symptoms using methods and techniques known in theart.

A common and important symptom of HD includes weight loss. This is animportant and widely accepted indicator of HD. In fact, unintendedweight loss frequently complicates the course of neurodegenerativedisorders and can contribute substantially to morbidity and mortality.Aziz, N. Ahmad, et al., JON 3062: 1872-1878 (January 2009). In fact,studies have also shown that weight loss in individuals with HDcorrelates with CAG repeat length. Aziz, N. A. et al., Neurology71:1506-1513 (2008). An increase in weight loss was observed inindividuals with high number of CAG repeats. Accordingly, the presentinvention relates to preventing or treating weight loss in an individualhaving HD and/or an increase number of CAG repeats by administeringestrogen, testosterone, precursors thereof or a combination thereof. Thedata show that administration of estrogen or testosterone preventsweight lost with HD and increases quality of life, as further describedherein. Weight loss can measured using a scale.

Seizures are also a common symptom of this form of HD. Brain tissueloss, memory loss and psychiatric symptoms including depression andanxiety are also observed. Mutant huntingtin is expressed throughout thebody and associated with abnormalities in peripheral tissues that aredirectly caused by such expression outside the brain. Theseabnormalities include muscle atrophy, cardiac failure, impaired glucosetolerance, osteoporosis and testicular atrophy.

The present invention involves selecting an individual with HD and/or anindividual with the mutant HD gene having greater than or equal to about36 CAG repeats. Selecting these individuals can be done in several ways.HD can be assessed clinically by assessing components of the UnifiedHuntington's Disease Rating Scale (UHDRS). Movement Disorders (vol.11:136-142, 1996) and Neurology (54:452-458, 2000). UHDRS is generallydivided into four components: motor performance, cognition, behavior andfunctional capacity. In particular, the following items are rated:Ocular Pursuit (horizontal, vertical), Saccade Initiation (horizontal,vertical), Saccade Velocity (horizontal, vertical), Dysarthria, TongueProtrusion, Finger Taps (right, left), Pronate/Supinate (right, left),Fist-Hand-Palm Sequence, Rigidity-arms (right, left), Bradykinesia,Maximal Dystonia (trunk, RUE, LUE, RLE, LLE), Maximal Chorea (Face, BOL,Trunk, RUE, LUE, LLE, RLE), Gait and Tandem Walking. Each item isassigned a grade from 0 to 4, and added up to quantify the progressionof HD symptoms. A rating of an abnormal gate to a severely abnormal gateindicates progression of HD in the individual. The present inventionincludes methods for preventing the UHDRS score from worsening, ascompared to the UHDRS prior to administration. Additionally, the presentinvention includes methods for maintaining the UHDRS score or reducingthe progression of HD as measured by UHDRS, as compared to an individualwith HD that is not subject to estrogen/testosterone administration.Brain tissue loss can be assessed with a head CT scan, head MRI scan, aPET scan, or other imaging techniques.

An individual can be genetically tested for the mutant HD gene and thenumber of CAG repeats, a test that indicates the presence or absence ofHD in an individual. CAG repeats can be assessed using e.g., a PCRassay. Mangiarini et al, 1996. Briefly, a sample from the individual canbe taken and assessed. The sample can be prepared using the methodsknown in the art. The DNA from the sample is prepared and southern blotsand hybridization assays can be done for this assessment. CAG repeatscan be sized by PCR using labeled primers. A polymerase (e.g., Taqpolymerase) can be used along with cycling conditions (e.g., 90″ @ 90°C., 25×(30″ @ 94° C., 30″ @ 65° C., 90″ @ 72° C.), 10′ @ 72° C.). PCRproducts can be sized using sequences and genotyping software. The PCRproducts from an HD affected individual can be analyzed by DNAsequencing techniques to determine the absolute number of CAG repeats.Additional methods known in the art can assess the presence of themutant HD gene, and/or the size or length of the CAG repeat.Furthermore, DNA tests for HD disease or other trinucleotide repeatdisorders can be done e.g., through targeted mutational analysis using anumber of laboratories that are commercially available. In anembodiment, an individual predisposed to HD having tested positive forthe expanded CAG repeats but prior to having symptoms can be selectedfor administration of estrogen, testosterone or both, as describedherein. Alternatively, one can assess the presence of the polyglutaminetract protein or nucleic acid using methods known in the art or laterdeveloped.

HD is one of several trinucleotide repeat disorders which are caused bythe length of a repeated section of a gene exceeding a normal range.Accordingly, in addition to treating HD, the present invention relatesto treating diseases characterized by a polyglutamin tract or CAGrepeats (e.g., greater than or equal to 36 CAG repeats), as describedherein. Such polyglutamine diseases include Dentatorubropallidoluysianatrophy (e.g., about 49-88 CAG repeats), Kennedy disease (e.g., about38-62 CAG repeats), Spinocerebellar ataxia Type 1 (e.g., about 49-88 CAGrepeats), Type 2 (e.g., about 33-77 CAG repeats), Type 3 (e.g., about55-86 CAG repeats), Type 6 (e.g., about 21-30CAG repeats), Type 7 (e.g.,about 38-120 CAG repeats), and Type 17 (e.g., about 47-63 CAG repeats).

Accordingly, an individual with a trinucleotide repeat disorder isselected with a DNA test that assesses the presence of an expanded CAGrepeats, or a test that assesses the presence of the poly Q tract.Individuals can also exhibit other symptoms described herein and theprogression of those symptoms, especially weight loss, indicates theprogression of the disease. Estrogen/testosterone administration,prevents one or more of the symptoms described herein from worsening, orreduces the severity of the symptom by at least about 5%, (e.g., 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%), as compared to the level ofseverity of the symptom just prior to administration, or as compared toan individual with the trinucleotide repeat disorder that is not subjectto estrogen/testosterone administration. In another embodiment, theestrogen/testosterone administration ameliorates or improves one or moresymptoms described herein by about 5%, (e.g., 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, or 90%).

Modes and Manner of Administration, Dosages

“Estrogen/testosterone administration” refers to administering one ormore of the following: estrogen, testosterone, a precursor thereof, or acombination thereof. As referred to herein, estrogen, a precursorthereof or a combination thereof is generally administered to a femaleindividual, and testosterone, a precursor thereof or a combinationthereof is administered to a male individual. Estrogen refers to a groupof steroidal based compounds involved in the estrous cycle. Examples ofsteroidal hormone compounds that can be used with the present inventioninclude testosterone (17beta-hydroxy-4-androsten-3-one), estrone (E1,3-hydroxy-1,3,5(10)-estratrien-17-one), estradiol (E2,1,3,5(10)-estratriene-3,17beta-diol), estriol (E3,1,3,5(10)-estratriene-3,16alpha,17beta-triol), and progesterone (e.g.,P4 (pregn-4-ene-3,20-dione)). Several of these compounds arecommercially available for other purposes, such as menopause orhypogonadism-testicular failure.

The composition of testosterone is as follows:

Similarly, the composition of estradiol includes:

Estrogen compounds now known or later discovered or developed can beused with the present invention.

Estradiol comes in the form of oral, transdermal, topical, injectable,and vaginal preparations. In certain aspects, the estradiol molecule canbe linked to an alkyl group at C3 position to facilitate theadministration. Examples of such compounds include estradiol acetate(oral and vaginal applications), estradiol cyprionate (injectable),ethinylestradiol (oral).

In particular, a form of estradiol that can be used with the presentinvention is estradiol cypioate, which marketed under the nameDEPO-Estradiol by Pfizer, Inc. DEPO-Estradiol contains an oil solubleester of estradiol 17β. The chemical name for estradiol cypionate isestradiol 17-cyclopentanepropionate. DEPO-Estradiol, in an embodiment isadministered intramuscularly by injection. In an embodiment, each mLcontains: 5 mg/mL, 5 mg estradiol cypionate, 5.4 mg chlorobutanolanhydrous (chloral derivative) added as preservative; in 913 mgcottonseed oil. The composition of estradiol cypioate is as follows:

Similarly, a form of testosterone includes DEPO-Testosterone and can beused with the administration in the present invention. In thisembodiment, DEPO-Testosterone is for intramuscular injection andcontains testosterone cypionate which is the oil-soluble 17(beta)-cyclopentylpropionate ester of the androgenic hormonetestosterone. Testosterone cypionate is a white or creamy whitecrystalline powder, odorless or nearly so and stable in air. It isinsoluble in water, freely soluble in alcohol, chloroform, dioxane,ether, and soluble in vegetable oils. The chemical name for testosteronecypionate is androst-4-en-3-one,17-(3-cyclopentyl-1-oxopropoxy)-,(17β)-. Its molecular formula is C27H40O3, and the molecular weight412.61. DEPO-Testosterone is commercially available in two amounts byPfizer, Inc.: 100 mg/mL and 200 mg/mL testosterone cypionate.

The structural formula is represented below:

The estrogen, testosterone, precursors thereof, or combinations thereofused in the present invention can be administered with or without acarrier. The terms “pharmaceutically acceptable carrier” or a “carrier”refer to any generally acceptable excipient or drug delivery compositionthat is relatively inert and non-toxic. Exemplary carriers includesterile water, salt solutions (such as Ringer's solution), alcohols,gelatin, talc, viscous paraffin, fatty acid esters,hydroxymethylcellulose, polyvinyl pyrolidone, calcium carbonate,carbohydrates (such as lactose, sucrose, dextrose, mannose, albumin,starch, cellulose, silica gel, polyethylene glycol (PEG), dried skimmilk, rice flour, magnesium stearate, and the like. Suitableformulations and additional carriers are described in Remington'sPharmaceutical Sciences, (17th Ed., Mack Pub. Co., Easton, Pa.). Suchpreparations can be sterilized and, if desired, mixed with auxiliaryagents, e.g., lubricants, preservatives, stabilizers, wetting agents,emulsifiers, salts for influencing osmotic pressure, buffers, coloring,preservatives and/or aromatic substances and the like which do notdeleteriously react with the active compounds. Typical preservatives caninclude; potassium sorbate, sodium metabisulfite, methyl paraben, propylparaben, thimerosal, chloral derivative, etc. The compositions can alsobe combined where desired with other substances used to treat HD diseaseor trinucleotide repeat disorders, e.g., tetrabenazin neurolepticsmedications, benzodiazepines, selective serotonin reuptake inhibitors,and mirtazapine. A carrier (e.g., a pharmaceutically acceptable carrier)is preferred, but not necessary to administer the compound.

The estrogen/testosterone compound can be a liquid solution, suspension,emulsion, tablet, pill, gel, capsule, sustained release formulation, orpowder. The method of administration can dictate how the compositionwill be formulated. For example, the composition can be formulated as asuppository, with traditional binders and carriers such astriglycerides. Oral formulation can include standard carriers such aspharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharine, cellulose, magnesium carbonate, etc.

The estrogen/testosterone compound used in the invention can beadministered intravenously, parenterally, intramuscularly (e.g.,DEPO-Estradiol, DEPO-Testosterone), subcutaneously, orally (e.g.,Cenestin, Estinyl, Estrace, Menest, Ogen, Premarin), nasally, topically(e.g., Estrace, Ogen, Ortho Dienestrol, Premarin, Androgel, Estrogel,Testim), by inhalation, by implant (temporarily, e.g., Estring,Femring), by injection (e.g., Testosterone Cypionate), by suppository(e.g., Vagifem) or transdermally (e.g., Androderm, Alora, Climara,Esclim, Estraderm, Vivelle, Vivelle-Dot). For testosteroneadministration, in an embodiment, DEPO-testosterone can be administeredin amounts of 40 mg/week intramuscularly. Androgel is a gel that isapplied topically twice per day in an amount of 5 gm/day or 35 gm perweek. For estrogen administration, in an embodiment, Estrogel isadministered once a day in an amount of 0.75 mg/day or 5.25 mg/week,whereas Vivelle-Dot is a patch that administered 0.025 mg/day or 0.175mg/week. The composition can be administered in a single dose or in morethan one dose over a period of time to confer the desired effect (e.g.,periodically, daily, weekly, monthly, yearly, etc.). In one embodiment,estrogen, a precursor thereof or metabolite thereof can be administeredin an amount between about.025 mg to about 10 mg. Such administration,according the present invention, brings the average levels of estrogenin an female individual with HD to homeostatic levels of between about100 pg/ml and about 185 pg/ml. In another embodiment, testosterone, aprecursor thereof or a metabolite there can be administered in an amountthat ranges between about 1 mg and about 35 g. Such administrationallows for homeostatic levels in male individuals with HD to be asfollows: free testosterone is between about 20 pg/ml and about 40 pg/ml,and total testosterone is between 300 ng/dL and about 900 ng/dL.

The actual effective amounts of compound or drug can vary according tothe specific composition being utilized, the mode of administration andthe age, weight and condition of the patient. For example, as usedherein, an effective amount of the drug is an amount which reduces oneor more HD symptoms. Dosages for a particular individual patient can bedetermined by one of ordinary skill in the art using conventionalconsiderations, (e.g. by means of an appropriate, conventionalpharmacological protocol).

For enteral or mucosal application (including via oral and nasalmucosa), particularly suitable are tablets, liquids, drops,suppositories or capsules. A syrup, elixir or the like can be usedwherein a sweetened vehicle is employed. Liposomes, microspheres, andmicrocapsules are available and can be used.

For parenteral application, particularly suitable are injectable,sterile solutions, preferably oily or aqueous solutions, as well assuspensions, emulsions, or implants, including suppositories. Inparticular, carriers for parenteral administration include aqueoussolutions of dextrose, saline, pure water, ethanol, glycerol, propyleneglycol, peanut oil, sesame oil, polyoxyethylene-polyoxypropylene blockpolymers, and the like. Ampules are convenient unit dosages.

The administration of a first estrogen/testosterone compound and asecond estrogen/testosterone compound or compound for treatingtrinucleotide repeat disorders can occur simultaneously or sequentiallyin time. The estrogen/testosterone compound can be administered before,after or at the same time as a second compound. Thus, the term“co-administration” is used herein to mean that the firstestrogen/testosterone compound and a second compound will beadministered at times to achieve a reduction in symptoms, or treatmentof HD or the trinucleotide repeat disorder. The methods of the presentinvention are not limited to the sequence in which the compounds areadministered; so long as the estrogen/testosterone compound isadministered close enough in time to produce the desired effect.

As hormones such as estrogen, testosterone, and their precursorsdecline, the protective effect of these hormones is lost and the HDdisease process advances undeterred. Specifically, for example, estrogendeclines by 50% in women by approximately 50 years of age and a further80% following menopause. In an aspect, testosterone's and estrogen's,and their precursors', effects on brain functions include, but are notnecessarily limited to, stimulation of the growth of the dendriticspines on spiny neurons and regulation of dopomenergic, serotonergic,adrenergic and glutamatergic functions. Testosterone, estrogen, andtheir precursors also increase the synthesis of certain monoamineneurotransmitters, inhibit their degradation, and interact withneurotrophins that stimulate neural growth and survival.

In an embodiment, a hormonal level of estrogen to be maintained in awomen ranges between about 100 pg/ml and about 185 pg/ml. With respectto men, free testosterone levels can be maintained in a range betweenabout 20 pg/ml and about 40 pg/ml, total testosterone levels can bemaintained in a range between 300 ng/dL and about 900 ng/dL, or both.The level of estradiol can be determined by taking an E2 test, and thelevel of testosterone can be determined by taking a serum testosteronetest, both of which are diagnostic blood tests that are commerciallyavailable. When the level of estrogen and/or testosterone falls belowthis level in an individual selected for HD (e.g., usually and optimallybefore recognition of any symptoms) or a trinucleotide repeat disorder,then an amount of estrogen and/or testosterone can be administered tokeep the levels in this range.

Maintaining estrogen and/or testosterone levels in individuals having HDor a trinucleotide repeat disorder can be done by administering theestgrogen/testosterone compound, as described herein, and can take theform of injection, transdermal patches and/or oral therapy.

Animal models of transgenic mice have been developed for the mutant HDgene and specifically it is known that these mice that carry thistransgenic gene containing the first exon of the mutant HD gene and havethe identical function as the human mutant HD gene. The brains of HDpatients and animal models of HD have specific abnormalities that arenegatively affected by deficiency of testosterone, estrogen, or theirprecursors. Abnormalities in monoamingeric neurotransmission areimplicated in the disease process. HD pathogenesis suggests that anearly excess of dopaminergic activity causes excitotoxic cell death thatis followed by a dopaminergic deficiency as neuronal death ensues.Estrogen's, testosterone's, and their precursors' knownanti-dopaminergic action in the striatum enables them to function earlyin the disease as neuroprotectants. A differential sensitivity toglutamate causes glutamate toxicity that, in turn, affects spiny neuronsin the striatum. Expression of the huntingtin polyglutamine expansion incells exposed to NMDA-type glutamate receptors causes increasedexcitotoxic cell death when compared to control cells. Estrogen,testosterone and/or their precursors act to suppress mRNA levels ofcertain NMDA-glutamate receptors and are thus able to protect neuronsagainst glutamate toxicity. Estrogen, testosterone, and/or theirprecursors mediate neuroprotection in neurons induced to undergoapoptosis. Apoptosis, or programmed cell death, is generally used bymulticellular organisms to eliminate unnecessary or dangerous cells.Apoptosis is normally important to the development of the brain andnervous system, the immune systems and various body tissues. However, inthe case of HD, apoptosis leads to excessive degeneration of nervecells. Apoptosis is induced by the mutant polyglutamine from thehuntingtin protein, causing an influx of Ca+ into a cell through theglutamate receptor. Genes are typically essential for Programmed CellDeath (PCD) and, by blocking the expressed gene products, such as the HDgene product that codes for the huntingtin protein, PCD is inhibited. Inthis case, estrogen, testosterone and/or their precursors bind to thepolyglutamine located on the end of the mutant huntingtin protein toprevent the protein from inducing cell death by preventing aggregateaccumulation on the nuclear membrane and in the nuclei of these neurons.The ability of estrogen, testosterone, and their precursors to bind tothe mutant huntingtin protein decreases as one ages because theproduction of estrogen, testosterone, and their precursors decreaseswith age. Since the length of the CAG polyglutamate repeat determinesthe level of hormone needed to render this protein ineffective incausing cell death, in an embodiment, it is an embodiment of the presentinvention, to begin treatment before ones natural reserves of hormoneare depleted with age. The extent of treatment with estrogen,testosterone, and/or their precursors will depend on a given individualbased on the length of the CAG glutamine repeat in that individual andthe stage of the disease. Elevated monoamine oxidase (MAO) activity inthe brain of HD patients is believed to be the basis for the depressivesymptoms so often characteristic of HD. Estrogen, testosterone, andtheir precursors inhibit MAO activity and have an antidepressant action.Estrogen's, testosterone's, and their precursors' ability to increasecerebral blood flow is believed to counter the decreased cerebral andcaudate blood flow associated with HD. Estrogen, testosterone, and theirprecursors serve as brain protectants in this process. Following is apreferred method for determining the rate of estrogen binding based onthe length of the CAG glutamine repeat. Comparable tests can then beused by physicians in a testing regimen in which hormone levels, such asestradiol levels, as well as the poly-glutamine repeat length of theindividual are measured to determine the optimum time to begin therapy.

EXEMPLIFICATION Example 1 In Vitro Studies Show that Estradiol Binds theMutant Form of Huntingtin Protein

Glycerol Bacterial stocks of three different CAG repeat lengthscontained within the first exon of Huntington and cloned into the PGexvector 2T were used to inoculate overnight cultures of 25 MI containing50 mglmL ampicillin. Pgex is a glutathione transferase fusion expressionsystem Amersham Cat#-27-4587-01-. This system allows for the efficientoverproduction of a target protein along with theglutathione-S-transferase as a fusion. These cultures were previouslygrown and DNA preparations of each were made. They were then sequencedusing dye terminator sequencing technique and run on an ABI 377 autosequencer. The results show a 23 CAG repeat contained within the firstexon of Huntingtin labeled Q23. A 47 CAG repeat contained within thefirst exon of Huntingtin labeled Q47 and a 63 CAG repeat containedwithin the first exon of Huntingtin labeled Q63. The bacterial cultureused to make this DNA prep was divided in half. Half of it was used forthe DNA prep (to specifically identify the number of CAG repeats usingdyterminator DNA sequencing) and the other half was used to make theglycerol stock used to inoculate these cultures to produceover-expression of the proteins containing three variables of the CAGrepeats. After overnight growth at 37 degrees, the cultures were diluted1:100 (e.g. 1.25 ML into 125 ml of LB containing 50 mg/ml of antibioticampicillin) and then allowed to continue to grow into middle log phase1.5 hours.

At this point, the bacterial cultures were made using 1.0 mMIsopropyl-b-dThiogalactopyranoside (IPTG) by diluting 100 mM IPTG 1:100(1.25 mL of 100 mM IPTG into 125 ml of growth culture representingapproximately 0.6 OD (optical density) at 590 nM using a common lightspectrophotometer). These three cultures were then allowed to growovernight (20 hrs.) at 30 degrees. These cultures were then centrifugedat 600 rpm for 30 minutes to collect the bacteria. The bacterial pelletswere then transferred to a 1.5 mL conical centrifuge tube and 1.0 mL ofice cold PBS (phosphate buffed saline) was added and the pelletsdissolved by vortexing for five minutes. This was then split into two1.5 mL conical tubes containing 700 μL each for all three bacterialpreps and then 350 μL of 3% N-lauyl sacosine was added to each of thesix tubes, mixed, and then each tube was sonicated for a total of 30seconds using a Fisher probe sonicator Model number F-50. Next the sixtubes were centrifuged in a microfuge top speed for 10 minutes.

The bacterial supernatant of a rich brown color was added directly tothe equivalent of 400 μL of Glutathione Sepharose 4B (strict affinityfor the glutathione portion of the fusion-target protein over produced)beads washed with ten bead volumes of ice cold PBS. The bacterialsupernatant, as a slurry with the Glutathione sepharose, was allowed togently rock for 30 minutes. After the binding was complete, the beadslurry was centrifuged at 200 rpm 20 seconds to pellet the bead and thenrepetitively washed five times with two bead volumes with ice cold PBS.After the last supernatant was removed and discarded, the beads weresubjected to elution with 10 mM reduced glutathione in 50 mM tris Ph 8.0by allowing incubation for 10 minutes under gentle shaking. These werethen centrifuged 30 seconds at 200 rpm and the elute removed and savedto a new tube. Next, 20 μl of protease inhibitor was added PMSF. Then 30μL of these were then subjected to polyacylamide gel electrophoresis(PAGE) 12% gels overnight at 90 volts constant voltage.

The next day the gels were stained with coomassie brilliant blue in 50%methanol and 10% acetic acid, for two hours at 50 degrees, and thenfurther destained in the same solution composition minus the coomassieblue for up to four hours. The resultant image shows purification of allthree mutant huntingtin protein bands.

These pure proteins were then used in the following binding protocol.Oestradiol-6-(0-carboxymethyl) oximino-(2-[125I]iodohistamine was usedas the estradiol source. Buffer B preferably comprises: 250 mM tris, 150mM NaCI, and a Dextran Charcoal solution comprising 0.05% Dextran coatedcharcoal in buffer B (sigma Cat#6197). About 0.5 ml of buffer B waspreferably used per reaction. A total of 5 ml of Buffer B was aliquotedinto a glass borosilicate tube (Fisher Cat#14-961-26) and 10 μL ofOestradiol (I125) was added containing approximately 2,000,000 cpm.About 0.5 ml of this solution was then distributed into eight labledglass tubes (cat #14-961-260). The entire tube #1's 500 μL was counted.Tube #2 through #8 were treated in the following manner:

TABLE 1 Tube #1 Total cpm in 0.5 mL of cocktail Tube #2 zero/0.5 mLcocktail Tube #3 Q23/0.5 mL cocktail + 50 μL Eluate 1 Tube #4 Q23/0.5 mLcocktail + 50 μL Eluate 1 Tube #5 Q47/0.5 mL cocktail + 50 μL Eluate 3Tube #6 Q47/0.5 mL cocktail + 50 μL Eluate 3 Tube #7 Q63/0.5 mLcocktail + 50 μL Eluate 5 Tube #8 Q63/0.5 mL cocktail + 50 μL Eluate 5Tubes #2-#8 were incubated for 1 hour at room temperature, placed on icefor ten minutes, and 200 μL of the Dextran charcoal solution in buffer Bwas added. These tubes were then placed on ice ten minutes and thencentrifuged 15 minutes at 2200 rpm in a Sorvall RC-3 refrigeratedcentrifuge with swinging bucket rotor (Sorvall HL-8) at 4 degrees. Theentire 700 μL supernatant was removed and counted using Genesysmulti-well gamma counter (Laboratories Technology, Inc.). The resultsare summarized below:

TABLE 2 Counts per minute (cpm): Total: 184,972 (I 125) estradiol inbuffer B (count entire tube) Zero: 27,388 Charcoal Filtered, countsupernatant only Sample HD Q23: 50,107 +HD Q23, Charcoal Filtered, CountSup. Sample HD Q23: 53,797 +HD Q23, Charcoal Filtered, Count Sup. SampleHD Q47: 47,454 +HD Q47, Charcoal Filtered, Count Sup. Sample HDQ47:44,767 +HDQ47, Charcoal Filtered, Count Sup. Sample HD Q63: 38,221 +HDQ63, Charcoal Filtered, Count Sup. Sample HD Q63: 38,040 +HD Q63,Charcoal Filtered, Count Sup.

The significance of these data is that an affinity for estradiol hasbeen demonstrated using the mutant form of huntingtin, the causativeprotein in Huntington's disease. It is therefore now applicable thatHuntington Disease patients and their at-risk family members should havetheir polyglutamine repeat length tested and if recognized, to start ona regimen of estradiol, or related hormone, replacement therapy as soonas their normal level of these hormones drop below physiologic levels.Physicians set up a testing regimen that determine both estradiol levelsas well as the poly-glutamine repeat length to determine the optimumtime to begin therapy. It is thereby shown that estradiol is thelimiting factor in the progression of Huntington's disease. Theestradiol level, or the serum level of other applicable hormones, inconjunction with the known CAG repeat length of affected individuals, istherefore predictive of the symptomatic onset of Huntington's Diseaseand thus will establish the optimum time in the individual's life tobegin administering the appropriate hormone therapy.

Example 2 In Vivo Treatment of HD Animals with Estrogen and TestosteroneAdministration Summary

HD is a neurodegenerative disorder caused by a CAG/polyglutamine repeatexpansion. Mice have been generated that are transgenic for the 5′-endof the human HD gene carrying CAG115-CAG160 repeat expansions. The mostsevere (CAG 158-160) repeat length was chosen for this study. Althoughthere was the possibility that any treatment would be ineffective atthis severity of disease state, even a minimal improvement in cognitiveand motor function is suggestive of a treatment.

In this study that Estrogen and/or Testosterone treatment of transgenicmice, even at this severe induction of disease, provide a significantimprovement in both cognitive decision making and motor function 14 days(16.6% improvement) after their untreated liter mates were unable toperform the trained task. Correlation of body and brain weights withthese improvements in treated animals reveals a treatment for HD.

Specific Aim:

The specific aim of this study is to determine if Estradiol/Testosteonetreatment of HD transgenic mice will delay or stop the onset of symptomsof HD.

Methods:

Thirty (30) hemizygous mice were tested to determine their ability totransverse a ½ inch square beam the distance of one meter. They wereinjected with estradiol cypionate (DEPO®-ESTRADIOL, Pfizer, Inc.) ortestosterone cypionate (DEPO®-TESTOSTERONE, Pfizer, Inc.) as an emulsionwith freunds incomplete adjuvant. Hormone tests of serum were performedto verify the blood levels of estradiol or testosterone. The mice(controls=no treatment) were tested against liter mates receiving either(E1 and E3 100 ug/ml weekly) or (E2 and E4 200 ug/ml injection weekly).The test will consist of Beam walking, (measuring coordination, slipfalls and distance in a measured time interval), brain weight and bodyweight. The testing will be performed twice weekly from week 5 throughweek 12 of age.

Two groups of mice, 15 female and 15 male mice will be utilized.

30 HD mice were obtained from the Jackson Laboratory, Bar Harbor, Me.The group consists of a 15 hemizygous female strain #2810 and 15hemizygous males strain #2810 (containing the transgene HDexon 1+159-161CAG repeat units). These mice were grouped as follows:

-   -   All mice received ear clips as identification, noting hair        color.    -   1-15 were males and 16-29 were females mouse #30 arrived dead.    -   Male mice: 1-4 were control animals receiving no treatment,        5,6,7,11 were grouped as E1 8,9,10 and 12 were grouped as E2.        13, 14 and 15 were grouped as T1, receiving 100 ug/ml        testosterone weekly.    -   Female mice: 16-19 were control animals receiving no treatment.        20,21,22,26 were grouped as E3, 23,24,25, 27 were grouped as E4.        28 and 29 were grouped as T2, receiving 100 ug/ml testosterone.        See table 3 for a summary.

TABLE 3 Male (M)/ Estradiol (E)/ Amount Numeral Female (F)Testosterone(T) received Designation E1 M E 100 ug/ml 5, 6, 7, 11 E2 M E200 ug/ml 8, 9, 10, 12 E3 F E 100 ug/ml 20, 21, 22, 26 E4 F E 200 ug/ml23, 24, 25, 27 T1 M T 100 ug/ml 13, 14, 15 T2 F T 100 ug/ml 28, 29Control M None None  1-4 Control F None None 16-19

Observations included behavioral testing and body weight selected tomeasure motor aspects of fore limb and hind limb coordination, balance,including raised beam. Blood specimens for estradiol assay using Elisamethodology was obtained during the study.

Estradiol/Testosterone Injection:

Stock solution of estradiol and testosterone were diluted with water andmixed with Freund's incomplete adjuvant. This was mixed continuously for1 minute and either 100 ul (E1 and E3) or 200 ul (E2 and E4) wereinjected subcutaneously. Estrogen or testosterone release was continuousover six days. Peak measured levels were obtained in 24 hr as shown inprevious studies. Animals in the E1 and E3 groups were targeted toreceive 100-300 pg/ml estrogen and groups E2 and E4 were targeted toreceive 400-800 pg/ml of estrogen. Animals in T1 and T2 grouping weretargeted to receive 100 pg/ml testosterone.

Behavioral Testing:

Behavioral testing included beam walking, brain weight and body weightassessment.

Beam Walking:

Utilizing the raised beam, mice were assessed for their ability totransverse a graded series of narrow beams to reach a safety platform.The beams consisted of long strips of wood with a 12 mm or 5 mm squarecross section. The beams were placed horizontally, 50 cm above eachbench surface, with one end mounted on a narrow support and the otherend attached to an enclosed box to which the animal could escape. Twoinches of dense foam was placed under the test area to protect theanimals from fall injury. Training took place in a 3 or 4 day period,where the mice were trained two to four times in a four day period usingthe 12 mm square beam. Once they were trained to walk the beam (i.e.,traversing the 12 mm square beam in less than 20 seconds), they receivedtwo consecutive trails on each of the square beam and each of the roundbeams, in each case progressing from the widest to the narrowest beam.Mice were afforded 150 seconds to traverse each beam. The number oftimes the mice slipped off of the beam was recorded for each trial. Themean score was used for each trial.

Body Weight: All animals were weighed on a scale weekly throughout thestudy. All weights were recorded.After euthanized, all brains of all mice were surgically removed andplaced in a solution of formaldehyde and weighed on a scale. The weightswere recorded.

Results:

A significant improvement was found in the ability of the estrogen andtestosterone treated animal to transverse the narrowest beam, a beamthat was ½ square inches. In motor skill, both hind limb and forelimbcoordination were statically improved over their liter mate controlsubjects as was shown by their ability to transverse the ½ in² beam. Themean average of CAG repeat length was 159.60, mice severely affectedwith HD. Two weeks after the control animals were unable to transverseeven one fourth of the beam, the estrogen/testosterone treated animalscontinued to be able to cross the ½ in² beam under the time allotted (a16.6% improvement over the course of this study). These observations ofcoordination and control of muscle movement further show the therapeuticeffects of estrogen and testosterone treatment of Huntington diseasedmice. Additionally, treated mice retained their body weightsignificantly better than non-treated mice. Table 4 shows the directcorrelation of hormone treated (estrogen and/or testosterone) mice andtheir body weight retention, which is directly related to their abilityto perform motor function tests.

Table 4 shows the direct correlation of hormone treatment with bodyweight in female mice. The average number of CAG repeat for all mice was159.60.

TABLE 4 Females Peak wt Last wt Change in % change Average % ES/TT ID #Weeks 8/9 Week 12 Grams in weight change pg/ml C, 16 18.4 g 13.5 g −4.9g −26.6% 21.7%  12.2 pg/ml C, 17 23.5 g 19.1 g −4.4 g −18.7% 21.7%  27.9pg/ml C, 18 20.5 g 17.7 g −2.8 g −13.6% 21.7%  17.1 pg/ml C, 19 22.1 g15.9 g −6.2 g −28.0% 21.7%  26.1 pg/ml E1, 20 25.0 g 18.2 g −6.8 g−27.2% 15.5%  49.8 pg/ml E1, 21 24.0 g* 21.7 g −2.3 g −9.0% 15.5% 261.5pg/ml E1, 22 19.9 g 16.9 g −3.0 g −15.0% 15.5% 371.9 pg/ml E1, 26 21.2 g18.9 g −2.3 g −10.8% 15.5% 258.3 pg/ml E2, 23 21.5 g* 20.5 g −1.0 g−4.6% 15.7% 902.9 pg/ml E2, 24 21.8 g 17.6 g −4.2 g −19.2% 15.7% 651.2pg/ml E2, 25 19.8 g 13.5 g −6.3 g −31.0% 15.7% Dead E2, 27 22.5 g 20.7 g−1.8 g −8.0% 15.7%  20.4 pg/ml T1, 28 27.6 g 17.4 g −10.2 g  −36.9%34.8% 647.0 pg/ml T1, 29 21.6 g 14.5 g −7.1 g −32.8% 34.8% —Table 5 shows a direct correlation between hormone treatment and bodyweight in male mice. All mice had an average CAG repeat of 159.60

TABLE 5 Males Peak wt Last wt Change in Average % ES/TT ID# Weeks 8/9Week 12 Grams % change Change pg/ml C, 1 25.9 g 18.4 g −7.5 g −28.9%27.2%  15.6 pg/ml C, 2 29.0 g 20.2 g −8.8 g −30.0% 27.2% — C, 3 23.4 g18.7 g −4.7 g −20.0% 27.2%  28.1 pg/ml C, 4 26.0 g 18.2 g −7.8 g −30.0%27.2%  14.1 pg/ml E3, 5 26.4 g 23.5 g −2.9 g −10.9% 15.9% 317.7 pg/mlE3, 6 26.2 g 19.6 g −6.6 g −25.0% 15.9% 443.5 pg/ml E3, 7 26.7 g 22.6 g−4.1 g −15.3% 15.9% 316.9 pg/ml E3, 11 24.8 g 21.7 g −3.1 g −12.5% 15.9%555.9 pg/ml E4, 8 26.2 g 22.9 g −3.3 g −12.5% 14.9% 586.6 pg/ml E4, 924.6 g 19.6 g −5.0 g −20.3% 14.9% — E4, 10 23.7 g 20.2 g −3.5 g −14.7%14.9% 388.1 pg/ml E4, 12 22.1 g 19.4 g −2.7 g −12.2% 14.9% 1,297 pg/mlT2, 13 23.4 g 17.1 g −6.3 g −26.9% 22.2% 468.1 pg.ml T2, 14 28.0 g 25.1g −2.9 g −10.3% 22.2% 937.7 pg/ml T2, 15 27.0 g 19.0 g −8.0 g −29.6%22.2% 1,494 pg/ml C = control animals, no treatment E1, E3 = 100 ul ESdose - target blood level - 100-300 pg/ml E2, E4 = 200 ul ES dose -target blood level - 400-800 pg/ml T1, T2 = 100 ul T dose - target bloodlevel - 50-100 pg/ml Control animals lost on average 21.7% of their peakbody weight (females) as compared to their treated liter mates, who lostan average of (E1) 15.5% and (E2) 15.7% weight loss. The male treatedanimal had similar findings. The control animals lost on average 27.2%of their peak body weight as compared to treated mice who hadexperienced a (E3) 15.9% and (E4) 14.9% weight loss. Weight loss is asignificant symptom of HD and usually attributed to mid onset of thediseased state.

Referring to the E1, Female ID #20, a sample from this particular mouseindicated that only about 49.8 pg/ml of estrogen was present, ascompared to other mice in the same group with 5 to 7 times that amount.For whatever reason, the mouse did not absorb the requisite or desiredamount of estrogen and accordingly, the mouse lost 27.2% of its bodyweight as compared to other members of the same group that only lost anbetween 4.6% and 15% of their body weight. This difference in bodyweight in light of the amount of estrogen absorbed is clear evidencethat estrogen administration/absorption directly correlates with weightloss prevention, one of the most important symptoms of HD.

The treated animals were better able to make decisions and coordinatedtheir forelimb and hind limb motor movements to cross the ½ in² beamwithout significant hesitation and within the time allotted. None of thecontrol (untreated) animals were able to coordinate their decisionmaking and motor function to cross the beam in the allotted time afterweek 10. Furthermore, if left for a longer duration of time (data notshown), the control animals just stayed on the start platform unable (orwilling) to move.

Table 6 shows the correlation between treated/untreated, CAG repeatlength and brain weight. On average the lower level (100-300 Pg/ml) ofestrogen treatment showed up to a 10% increase in brain weight.

TABLE 6 Animal ID# Brain weight at week 12 CAG repeat length C, 1 0.36 g163 C, 2 — 159 C, 3 0.36 g 160 C, 4 0.23 g 161 E3, 5 0.40 g 159 E3, 60.37 g 159 E3, 7 0.37 g 160 E4, 8 0.39 g 158 E4, 9 — 159 E4,10 0.34 g159 E3,11 0.40 g 158 E4,12 0.35 g 160 T2,13 0.31 g 159 T2,14 0.37 g 159T2,15 0.33 g 161 C,16 0.32 g 160 C,17 0.35 g 160 C,18 0.33 g 158 C,190.33 g 160 E1,20 0.33 g 159 E1,21 0.36 g 161 E1,22 0.34 g 159 E2,23 0.33g 158 E2,24 0.37 g 162 E2,25 — 162 E1,26 0.30 g 158 E2,27 0.37 g 159T1,28 0.31 g 159 T1,29 — 160

FIG. 1 shows the combined data of six mice, 3 controls and both estrogenlevels in 2 male and 1 female mice. All control animals were unable tocomplete the task of traversing the ½ in. beam in 150 seconds, whereasall of the treated animals shown were able to complete the traversing ofthe beam in the allotted time shown.

FIGS. 2A, 2B, 3A, 3B, and 4A, 4B show the data comparing one controlmouse to one treated animal. In all cases shown the treated animalsdemonstrated a significant increase in their ability to cross the beamwithin the time allotment. Additionally the data show the ability of thetreated animals to completely transverse the beam as measured bydistance. The untreated control animals were unable to cross even theshortest distance after week 10.

Although specific features of the invention are described in connectionwith some of the preferred methods and not others, this is forconvenience only as some feature may be combined with any or all of theother features in accordance with the invention. Other embodiments willoccur to those skilled in the art and are within the following claims.

This application relates to U.S. application Ser. No. 12/961,250, titled“Methods for Inhibiting the Development of Huntington's Disease”, filedDec. 6, 2010, by James A. Carnazza; U.S. application Ser. No.12/719,361, titled “Methods for Testing for and Inhibiting theDevelopment of Huntington's Disease”, filed Mar. 8, 2010, now abandoned,by James A. Carnazza; U.S. application Ser. No. 11/627,978, titled“Methods for Testing for the Development of Huntington's Disease”, filedJan. 28, 2007, issued as U.S. Pat. No. 7,674,592 on Mar. 9, 2010, byJames A. Carnazza; U.S. application Ser. No. 10/654,850, titled “Methodsfor Testing for and Inhibiting the Development of Huntington's Disease”,filed Apr. 9, 2003, now abandoned, by James A. Carnazza; U.S.Provisional Application No. 60/443,397, titled “Method for Inhibitingthe Development of Huntington's Disease”, filed Jan. 29, 2003, by JamesA. Carnazza; and U.S. Provisional Application 60/408,184, titled “Methodfor Inhibiting the Development of Huntington's Disease in Individualswith an Expanded CAG Trinucleotide in the Gene that Codes for theHuntington Protein”, filed Apr. 9, 2002, by James A. Carnazza.

The relevant teachings of all the references, patents and/or patentapplications cited herein are incorporated herein by reference in theirentirety.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed is:
 1. A method of treating an individual having atrinucleotide repeat disorder, the method comprises: a. selecting theindividual having the trinucleotide repeat disorder; and b.administering to the individual an amount of 17β-estradiol ortestosterone, wherein the amount of administration to an individualranges from about 0.2 mg to about 10 mg of 17β-estradiol for a femaleindividual or about 1 mg to about 35 g of testosterone for a maleindividual; wherein one or more symptoms associated with thetrinucleotide repeat disorder in the individual are reduced, as comparedto the same in an individual with the trinucleotide repeat disorder notsubjected to step b.
 2. The method of claim 1, wherein the individualhaving the trinucleotide repeat disorder has between about 36 and about120 repeats.
 3. The method of claim 1, wherein a trinucleotide repeatdisorder comprises Huntington's Disease (HD), Dentatorubropallidoluysianatrophy, Kennedy disease, Spinocerebellar ataxia (SA) Type 1, SA Type 2,SA Type 3, SA Type 6, SA Type 7, and SA Type
 17. 4. The method of claim1, further comprising measuring levels of 17β-estradiol in the femaleindividual or measuring levels of testosterone in the male individual.5. The method of claim 1, wherein the individual has greater than 40repeats.
 6. The method of claim 1, wherein one or more of the symptomsare reduced by between about 5% and about 100%.
 7. A method of reducingone or more symptoms associated with a trinucleotide repeat disorder inan individual, wherein the method comprises: a. selecting the individualhaving the trinucleotide repeat disorder; and b. administering to theindividual an amount of 17β-estradiol or testosterone, wherein theamount of administration to an individual ranges from about 0.2 mg toabout 10 mg of 17β-estradiol for a female individual or about 1 mg toabout 35 g of testosterone for a male individual; wherein one or moresymptoms associated with the trinucleotide repeat disorder in theindividual are reduced, as compared to the same in an individual withthe trinucleotide repeat disorder not subjected to step b.
 8. The methodof claim 7, wherein one or more symptoms are reduced by an amountbetween about 5% and about 100%.
 9. A method of treating an individualhaving Huntington's Disease (HD) or an individual having an HD genehaving a number of CAG repeats greater than or equal to about 36; themethod comprising: a. selecting the individual having HD; and b.administering to the individual an amount of estrogen or testosterone,wherein the amount of administration to an individual ranges from about0.2 mg to about 10 mg of estrogen for a female individual or about 1 mgto about 35 g of testosterone for a male individual; wherein one or moresymptoms associated with HD in the individual are reduced, as comparedto the same in an individual with HD not subjected to step b.
 10. Themethod of claim 9, wherein the symptoms of HD comprise weight loss, lossof fine motor function, loss of gross motor function, a loss incognitive function, chorea, loss of brain tissue, or a combinationthereof.
 11. The method of claim 9, wherein one or more symptoms arereduced by an amount between about 5% and about 100%.
 12. The method ofclaim 9, further comprising measuring levels of estrogen in the femaleindividual.
 13. The method of claim 9, wherein the female individual hasgreater than 40 CAG repeats.
 14. The method of claim 9, wherein thefemale individual has between about 36 and about 120 CAG repeats. 15.The method of claim 9, wherein one or more symptoms of HD is reduced,wherein the symptoms of HD comprise weight loss, loss of fine motorfunction, loss of gross motor function, a loss in cognitive function,chorea, loss of brain tissue, or a combination thereof.
 16. The methodof claim 15, wherein one or more of the symptoms are reduced by betweenabout 5% and about 100%.
 17. The method of claim 15, wherein the UnifiedHuntington's Disease Rating Scale (UHDRS) score remains about the sameor does not increase.
 18. The method of claim 9, wherein selecting afemale individual having HD comprises assessing the presence or absenceof a mutant HD gene having between about 36 and 120 CAG repeats.